Cloning and analysis of promoter-active fragments from Corynebacterium glutamicum 10147
谷氨酸棒杆菌10147基因组中启动子活性片段的克隆与分析

Abstract: Objective] To clone promoter-active fragments from Corynebacterium glutamicum for further construction of expression vectors. Methods] Random Sau3A I digested fragments of C. glutamicum 10147 chromosome were shot-gun cloned into the promoter-probe vector pAKC6 and promoter activity of the inserted fragments was selected by chloramphenicol resistance of transformed C. glutamicum cells. Results] Thirty promoter-carrying fragments were isolated. Three C. glutamicum clones harboring pAKC6 with promoter fragments displayed chloramphenicol acetyltransferase (CAT) activity of more than 24 U/mg. The fragment F57 led to the highest CAT activity of 32.50 U/mg, even more than that produced by the promoter Ptrc, 26.33 U/mg. Conclusion] The strength of promoter on fragments F21, F54 and F57 is as strong as promoter Ptrc in C. glutamicum. These fragments can be used to construct expression vector.